Maize-targeted mutagenesis: A knockout resource for maize
Maize-targeted mutagenesis: A knockout resource for maize
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DOI：
10.1073/pnas.1831119100
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发表时间：
2003-09-30
影响因子：
11.1
通讯作者：
Martienssen, R
中科院分区：
文献类型：
--
作者：
May, BP;Liu, H;Martienssen, R
我们描述了一种用于玉米定点转座子诱变的有效系统。利用Robertson's Mutator(Mu)花粉亲本产生了总共43,776株F-1植株，并进行自花授粉，建立了转座子诱变种子库。每 F-1 植物新种子突变体的频率在 10(-4) 和 10(-5) 之间。作为对玉米群体的一项服务，玉米定向诱变通过使用 PCR 从该文库中选择感兴趣基因的插入。谱系、敲除、序列、表型和其他信息存储在强大的交互式数据库（玉米定向诱变数据库）中，可以分析整个群体并处理敲除请求。通过抑制大多数 F-1 植物中的 Mu 活性，我们试图减少可能导致从混合组织中选择的假阳性的体细胞插入。然而，通过监测 F-2 中剩余的 Mu 活性，我们证明种子表型依赖于它，并且在似乎缺乏它的品系中出现假阳性。我们得出的结论是，该群体中出现的所有突变中有一半以上因失去 Mu 活性而受到抑制。这些结果对近亲繁殖的表观遗传模型和功能基因组学具有重要意义。
We describe an efficient system for site-selected transposon mutagenesis in maize. A total of 43,776 F-1 plants were generated by using Robertson's Mutator(Mu) pollen parents and self-pollinated to establish a library of transposon-mutagenized seed. The frequency of new seed mutants was between 10(-4) and 10(-5) per F-1 plant. As a service to the maize community, maize-targeted mutagenesis selects insertions in genes of interest from this library by using the PCR. Pedigree, knockout, sequence, phenotype, and other information is stored in a powerful interactive database (maize-targeted mutagenesis database) that enables analysis of the entire population and the handling of knockout requests. By inhibiting Mu activity in most F-1 plants, we sought to reduce somatic insertions that may cause false positives selected from pooled tissue. By monitoring the remaining Mu activity in the F-2, however, we demonstrate that seed phenotypes depend on it, and false positives occur in lines that appear to lack it. We conclude that more than half of all mutations arising in this population are suppressed on losing Mu activity. These results have implications for epigenetic models of inbreeding and for functional genomics.